Innovare International Journal of Pharmacy and Pharmaceutical Sciences

Academic Sciences
ISSN- 0975-1491 Vol 7, Issue 11, 2015

Original Article
OCULAR TOLERANCE AND IN-VITRO RELEASE OF CHLORAMPHENICOL IN PROSPECTIVE EYE
OINTMENT BASES

AYOBAMI O. OYEDELEa*, OKIKIOLU O. JOHNb, HANNAH O. OGUNGBEMIa, SAMUEL O. OLATEJUc

aDepartmentof Pharmaceutics, Faculty of Pharmacy, Obafemi Awolowo University, Ile-Ife, Nigeria, bDepartment of Clinical Pharmacy and
Pharmacy Administration, Faculty of Pharmacy, Obafemi Awolowo University, Ile-Ife, Nigeria, cDepartment of Surgery, College of Health
Sciences, Obafemi Awolowo University, Ile-Ife, Nigeria
Email: aoyedele@oauife.edu.ng
Received: 14 Aug 2015 Revised and Accepted: 03 Oct 2015
ABSTRACT
Objective: Chloramphenicol is poorly released from official eye ointment (EO) base. This study investigated alternative promising bases, enhanced
drug release with surfactant and evaluated ocular tolerance of prototypes.
Methods: Chloramphenicol eye ointment (1 %w/w) was prepared by levigation in five bases: EO, simple ointment (SO), hydrous wool fat (HW),
hydrous sheabutter (HS), and neat sheabutter (NS). In-vitro drug release of these and EO and SO formulations containing graded concentrations
(0.1–10.0 %w/v) of surfactant (polysorbate 80 or propylene glycol) were studied over 3 h by dialysis technique, and the release kinetics assessed.
Water-absorption capacity (WAC) and melting temperature of the bases was determined for probable influence on drug release. Ocular tolerance of
the formulations, bases and surfactants was evaluated by in vivo irritancy test on albino rabbits.
Results: Total drug released from test formulations differed with base used: NS 12, HS 11, SO 7, HW 5, and EO 4 %. The Higuchi release kinetics was
determined. HS and NS demonstrated highest release extent and rate; EO and HW exhibited poor release. WAC and softening temperature of bases showed
some correlation with drug release propensity. Propylene glycol in EO and polysorbate 80 in SO formulations showed 3-fold and 2-fold enhanced drug
release, respectively. All items for ocular toxicity test gave scores indicating practically no irritation, except neat HW that was mildly irritant.
Conclusion: HS and NS proved best alternatives to EO. Propylene glycol (10 %) in the EO formulation was 3-fold drug-release enhanced. All
prototype formulations were non-irritant to eye tissue.
Keywords: Chloramphenicol, Ointment, Base, Surfactant, In-vitro Release, Ocular tolerance.

INTRODUCTION vehicle for eye ointment medication is required to be non-irritant to

Chloramphenicol is a broad-spectrum antibiotic often applied
topically to conjunctiva for treatment of infections caused by
susceptible microorganisms. It is the drug of choice in ocular
infections because of its high transcorneal penetration and broad
spectrum activity [1] and is administered as ophthalmic drops or
ointment. The ointment offers an advantage of longer contact time
over the solution drug form that suffers rapid and extensive pre-
corneal loss due to high lachrymal fluid turn over and drainage rate
[2]. The potency of a topical medication is markedly influenced by its
vehicle, e. g. an ointment base [3], and the corneal absorption of the
administered drug may be enhanced by the presence of a surfactant
[4]. Non-ionic surfactants (such as polysorbate 80) are found to be
less toxic than anionic and cationic surfactants in predictive ocular
irritancy studies [5] and are uniquely suited for topical ocular drug
delivery through their potential ability to increase bioavailability by
increasing drug solubility, prolonging pre-corneal retention, and
enhancing permeability [6]. Polysorbate 80 (aqueous solution) has
been used to produce nanoparticles for ocular delivery of
hydrocortisone [7] and as a surfactant component of ocular
indomethacin microemulsion [8]. Propylene glycol is a water-
diffusive liquid with surfactant property and useful as cosolvent and
skin penetration enhancer for poorly soluble compounds in topical
medicinal products [9]. When tested on rabbit eye, neat (undiluted)
propylene glycol showed some measure of ocular irritation [10].
Also, occupational exposure to propylene glycol vapour has been
reported to cause ocular irritation in human subjects [11]. However,
dilutions of propylene glycol (at use concentrations) are generally
non-toxic [12]. In vivo Draize eye test in rabbit [13] is the
international standard assay for acute ocular toxicity [14].
Eye ointment basis BP (EO) is a British Pharmacopoeial (standard)
eye ointment vehicle generally recommended for use in eye
ointment preparations [15]. But preliminary study in our laboratory
suggested that chloramphenicol is poorly released from EO. Also, a
the conjunctiva [15]. Yet, no information is found in literature on
irritancy appraisal or ocular tolerance of EO, which should
authenticate its suitability and general acceptability for ophthalmic
use. Although the main components of EO (liquid and soft paraffin)
are common pharmacopoeial ointment vehicle components which,
in earlier ocular irritation studies, had been used as plain diluent or
medium [16, 17], the lack of any literature data indicative of the
ocular tolerance of EO or its components, even in later report on
predictive eye irritation potentials of several substances in current
use [18], is quite undesirable.
Other established (pharmacopoeial) ointment bases were having
similar components (paraffin, lanolin) to EO namely, simple
ointment BP (SO) and hydrous wool fat BP (HW) are, however, for
dermatologic use. These and sheabutter bases (hydrous sheabutter
[HS] and neat sheabutter [NS]) are investigated in this study, being
considered as prospectively useful alternative ophthalmic ointment
vehicles for chloramphenicol. Sheabutter, extracted and purified
from Vitellaria paradoxa C. F. Gaertn (Sapotaceae) nuts, has been
earlier studied for use as a dermatologic ointment vehicle and its
utility found comparable to that of established bases [19, 20].
However, for lack of satisfactory, unequivocal information on its
safety for human use, sheabutter was listed by the European
Commission 2006 among substances not exempted from
registration [21].
A valuable dermatologic vehicle, nonetheless, holds promise for
advantageous ophthalmic application if it is innocuous on the eye tissue.
Ocular tolerance testing of promising dermal ointment vehicles could
lead to expansion of their utilities. Hence it was adjudged important to
ascertain, by testing, the ocular toxicity potential of EO and of the
propitious dermatologic bases: SO, HW, HS and NS as necessary premise
for recommendation or proscription of their ophthalmic use. Therefore
the objectives of this study were to determine ocular irritation potential
of the bases and of chloramphenicol eye ointment prepared with each
 Oyedele et al.
base in the absence and presence of polysorbate 80 or propylene glycol
as drug-delivery enhancement surfactant; and to evaluate drug release
propensity of the formulations.
MATERIALS AND METHODS
Materials
Chloramphenicol in powder, anhydrous (SIGMA); cellulose dialyzer
tubing (SIGMA); propylene glycol (May and Baker, England);
cetostearyl alcohol, lanolin anhydrous (British Drug Houses,
England); polysorbate 80, hard paraffin, liquid paraffin, white soft
paraffin (William Ransom & Sons. England); neat sheabutter, freshly
processed from shea nuts (National Center for Agricultural
Mechanization, NCAM, Ilorin Nigeria).
Table 1: Composition of ointment bases
Name of base/Ingredient concentrations (%w/w)
Ingredients Eye ointment basis BP Simple ointment BP
Soft paraffin 80 85
Wool fat 10 5 75
Liquid paraffin 10 – –
Hard paraffin – 5
Cetostearyl alcohol – 5
Water (purified) – –
Neat sheabutter – –
Key:–Ingredient not contained in base
Determination of softening/melting point and water-
absorption capacity of ointment bases
Softening and melting temperatures of the bland bases were
determined using the method for suppository bases as described by
Adebayo and Akala [23]. The temperature at which the base sample
began to liquefy was defined as the softening point, while the
temperature of complete liquefaction was the melting point. The
mean and standard deviation (SD) of four determinations was
recorded. The possible influence of water-absorption capacity
(WAC) of ointment base on drug release propensity of the composite
(medicated) ointment formulation was assessed. Mean WAC of each
base was determined from quadruplicate tests at the ambient
temperature, using the pharmacopoeial method described for
hydrous wool fat [15].
Preparation of medicated ointment
Chloramphenicol eye ointment (1 %w/w) samples, 20 g each, were
prepared at the ambient temperature with each of the five ointment
bases by levigation method and aseptic processing, for subsequent
eye irritation and drug release testing. The medicated ointment
samples in EO and SO containing either propylene glycol or
polysorbate 80 (surfactant) at graded concentrations (0.1, 0.5, 1.0,
5.0 or 10.0 % w/v) were also prepared for drug-release
enhancement testing. The requisite amounts of the drug (≤ 90 μm
particle size) and surfactant were incorporated into each base,
respectively. Each preparation was aseptically packaged in sterile
disposable 2 or 5-ml capacity hypodermic syringes from which to be
dispensed at the point of use, and kept for a minimum of 24 h at the
ambient temperature before testing, to permit equilibration with the
environment.
In-vitro drug release studies
The release rate of chloramphenicol from each medicated ointment
sample was determined using an in-vitro dialysis technique
described earlier [19]. The release compartment was a 4 g
medicated ointment sample enclosed in 8 cm long cellulose dialyzer
tube previously hydrated in water for 24 h at room temperature,
suspended vertically on the rotary shaft of an Erweka dissolution
apparatus (Heusenstamm Kr. Offenbach, Germany) and set to
revolve at 50 rpm. The dissolution medium was a phosphate buffer
solution (900 ml, pH 7.2) maintained at 37±0.5 °C, simulating
lachrymal fluid pH [24]. Samples of the dissolution-medium (5 ml
Int J Pharm Pharm Sci, Vol 7, Issue 11, 306-311
Methods
Preparation of bases for chloramphenicol eye ointment
Five ointment bases (EO, SO, HW, HS, and NS; table 1) were
prepared. The EO, SO, and HW compounded according to
compendium guidelines [15], and HS produced from NS were
prepared sterile. EO and SO were dry-heat sterilized (BP procedure)
while HW, HS and NS were oven-melted (≤ 60 °C) and sterilized by
membrane filtration aided by suction pump. Freshly sterilized HS
and NS base samples having polymorphic property were kept
(undisturbed) at the ambient temperature (28±2 °C) for 5 d before
use, to allow full restoration and stability of their physical
characteristics and semisolid consistency [22].
Hydrous wool fat BP Hydrous sheabutter
– –
–
–
– –
– –
25 18
– 82
each) were withdrawn at specified time intervals for 180 min and
assayed for chloramphenicol. The volume of dissolution medium
was kept constant by replacing withdrawn volumes with equal
amount of fresh medium maintained at the same temperature. The
amount of drug in withdrawn samples was analyzed
spectrophotometrically at 278 nm (JENWAY 6305, UK). A calibration
graph was generated from a concentration range of the drug (0.5 to
4.0 mg/ml) prepared in pH 7.2 phosphate buffer solution which
showed a λmax of 1.239 UV absorbance at 278 nm wavelength, from
which the amount of drug in solution at each sampling time was
determined.
In vivo study
Animal ethics and experimental conditions
The study was approved by the Health Research and Ethics
Committee of Obafemi Awolowo University (OAU) Ile-Ife Nigeria
(Ref. no. OAUTHC/CS/232/Vol. V/268), and executed according to
the Test Guidelines no. 405 of the Organization for Economic
Cooperation and Development (OECD) in adherence to the
Association for Research in Vision and Ophthalmology (ARVO)
Statement for the use of animals in ophthalmic and vision research.
Healthy, New Zealand White albino rabbits (0.7–2.0 kg) of both
sexes without pre-existing ocular irritation was selected for the
study. The females were nulliparous and non-pregnant. The animals
were nurtured in the animal house of the Faculty of Pharmacy, OAU
Ile-Ife; housed in 56 × 56 × 46 cm well ventilated stainless steel
suspended cages, and were provided with standard feed from the
OAU Biological Gardens Unit, occasionally supplemented with herb
(Tridax procumbens Linn), and water ad libitum. The animal room
was maintained at temperature 26-30 °C, humidity 50-80 %, and
light/dark cycle 12h/12h conditions. Prior to dose initiation, the
rabbits were acclimatized to laboratory conditions for 21 d.
Primary eye irritation test on rabbits
The potential of each test item to cause irritation from a single
ocular instillation in a group of 3 rabbits as experimental bio-model
was assessed as recommended by the OECD, to determine the ocular
tolerance profile. The test items were: 0.1 g of each ointment base
and of the medicated ointment prepared in each base, and 100 µl of
each surfactant solution (10 %w/v) in purified water. Three animals
were used per test substance, which was instilled into the
conjunctival sac of the right (test) eye of each rabbit by gently
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pulling the lower lid away from the eyeball. The upper and lower RESULTS
lids were then gently held together for ~ 1 s to avoid loss of the test
substance. The left eye of each animal remained untreated as Amounts and profile of chloramphenicol release from ointment
control. Following treatment, ocular irritation was evaluated bases
macroscopically using a high-intensity white light (Mag Lite) at 1, The total amount of drug released from the chloramphenicol eye
24, 48, and 72 h post-instillation. ointment formulations over 3 h testing period depended on the
To minimize the number of animals used, any test animal showing no ointment base used. Over the period, the neat bases: NS released 12 %,
untoward response to applied treatment was reused once only, after a HS 11 %, SO 7 %, HW 5 %, and EO 4 % of the drug (fig. 1). Also, while
14-day rest/re-acclimatization period under the laboratory conditions. NS and HS gave gradual increase of the drug released over 3 h, the SO,
Twenty-four animals in all (10 males, 14 females) were used in the HW and EO formulations exhibited a drug release peak and no further
study. Individual eye irritation scores were recorded for each animal. appreciable release beyond 60, 120 and 120 min, respectively.
Absence of corneal damage at 24 h was verified by fluorescein dye
evaluation. Ocular lesions were scored according to the Scale for
Scoring Ocular Lesions [25]. Any other observed lesions were noted in
addition to observations of the cornea, iris and conjunctivae. The
rabbits were also monitored at least once daily for signs of gross
toxicity or behavioral changes during the test period. The average
score for all 3 rabbits per group at each scoring point was calculated.
For evaluation of the overall eye irritation scores, the time interval
with the highest mean score (Maximum Mean Total Score, MMTS) for
all rabbits in the group was used to classify the test substance in
accordance with the scoring system of Kay and Calandra [26].
Data analysis and statistics
The extent of drug release was assessed from the total amount of drug
present in the dissolution medium at the end of 180 min. The drug
release kinetics applicable for the ointment samples was evaluated by
analyzing with three mathematical models: zero-order kinetics (Q vs
t), diffusion-controlled (or Higuchi) model (Q vs square-root of t), and
first order kinetics (log[Qo–Q] vs t), where Q is the amount of drug Fig. 1: Release profile of chloramphenicol (1 %w/w) eye
released at time 't' and Qo is the initial amount of the drug. The model ointment in different bases
that consistently produced the highest correlation for the ointment
preparations was used for assessment of drug release rates, and the
slope obtained from linear regression analysis of the plot was For determination of chloramphenicol release kinetics, consistent
determined as the drug release rate constant. The results expressed as highest correlation of linear plots occurred when the amounts of
mean±SE (standard error of the mean percent drug-released values of drug released were plotted against the square root of time in
pooled data) were generated from triplicate determinations for each accordance with the Higuchi kinetics. The correlation coefficient
ointment formulation. The data were subjected to t tests, analysis of values obtained for most data of other kinetic models (zero and first
variance (ANOVA) and F-test to determine significance of mean drug- order) were consistently lower (table 2), implying preponderance of
released value differences. the diffusion-controlled release mechanism for the drug.

Table 2: Diffusion-controlled rates and correlation coefficients of chloramphenicol release mechanism profiles from ointment bases
Ointment base Diffusion-controlled model First-order model Zero-order model correlation coefficient
Release rate constant Correlation correlation coefficient
(mg. min–½) coefficient
Eye ointment basis 0.07 0.88 0.88 0.88
BP
Simple ointment 0.27 0.87 0.72 0.71
BP
Hydrous wool fat 0.17 0.98 0.94 0.94
BP
Hydrous 0.34 0.98 0.94 0.94
sheabutter
Neat sheabutter 0.28 0.94 0.93 0.92
The chloramphenicol release rate constant values in the ointment bases ranked in the order: HS (0.34)>NS (0.28)>SO (0.27)>HW (0.17)>EO (0.07
mg. min–½). Thus, HS and NS demonstrated greater release potential for chloramphenicol than the pharmacopoeial bases (SO, HW and EO). When
analyzed by ANOVA test, the release profile of chloramphenicol in the different ointment groups overall (i.e. by F test) differed significantly from
one another (*P>0.05). However, t test comparisons showed that the release data of HS compared to NS formulations was not significantly different
(*P<0.05).

Effect of water-absorption capacity and softening temperature
of base on drug release
The WAC and softening of the bases apparently influenced their
drug release propensity. With exception of HW, the bases (HS and
NS) softening at ~37 °C and having relatively lower WAC values
showed greater propensity for release of chloramphenicol than the
bases (EO and SO) with higher softening temperature and WAC
values (table 3). 37 °C is mammalian mean body temperature and
the in-vitro test condition. Inclusion of a surfactant as a component
of chloramphenicol eye ointment enhanced drug release from the EO
and SO formulations selectively. Thus 10 % propylene glycol
increased the amount of drug released in EO ointment from 4 %
(over 3 h) in an absence of the surfactant (fig. 1) to 13.5 % (in ≥1.5
h) when the surfactant was present (table 4).
This represents more than three-fold enhancement of the drug
release capacity of the base. Similarly, 10 % polysorbate 80 raised
the drug released in SO ointment from 7 % (over 3 h) in absence of
the surfactant to 18.9 % (in≥2 h) when the surfactant was present,

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Int J Pharm Pharm Sci, Vol 7, Issue 11, 306-311

which represents more than two-fold enhancement of the drug glycol and polysorbate 80 (surfactant) in SO and EO ointments,
liberating potential of the base. However, the presence of propylene respectively, did not appreciably improve drug release (table 4).

Table 3: Melting temperatures and water absorption capacity of ointment bases

Ointment base Heat susceptibility Water-absorption capacity (ml/10 g)†
Softening temperature ( °C) Melting temperature ( °C)
Eye ointment basis BP 38.5±0.5 44.0±0.5 35.0 (0.4)
Simple ointment BP 40.0±0.5 46.0±0.5 36.5 (0.4)
Hydrous wool fat BP 37.0±1.0 43.0±2.0 20.5 (0.4)
Hydrous sheabutter 37.5±2.0 42.5±1.5 10.5 (0.2)
Neat sheabutter 37.0±0.5 38.5±0.5 19.0 (0.2)
Key:-Mean values; with SD values in parenthesis

Table 4: Effect of surfactant on release of chloramphenicol from ointments prepared with Eye ointment BP or Simple ointment BP base
Time (h) ncentration of surfactant (%w/v) Base: Eye ointment BP Base: Simple ointment BP
Surfactant/Mean percent drug releaseda Surfactant/Mean percent drug releaseda
Propylene glycol Polysorbate 80 Propylene glycol Polysorbate 80
0b 2.8 (1.1) 2.8 (1.1) 4.1 (0.4) 4.1 (0.4)
0.1 5.7 (2.0) 0.7 (0.5) 0.9 (1.0) 8.5 (6.0)
0.5 0.5 4.1 (2.0) 2.5 (5.1) 1.9 (2.0) 7.7 (5.0)
1 3.9 (1.1) 4.7 (2.0) 2.2 (2.3) 8.5 (2.0)
5 4.8 (0.8) 3.2 (2.0) 3.0 (1.5) 5.1 (1.5)
10 8.0 (1.0) 4.5 (2.0) 8.1 (1.0) 10.6 (1.0)
0b 2.8 (1.1) 2.8 (1.1) 6.8 (0.4) 6.8 (0.4)
0.1 5.7 (2.0) 0.8 (0.5) 2.2 (1.0) 11.2 (6.0)
1.0 0.5 5.0 (2.0) 4.8 (5.1) 3.4 (2.0) 9.4 (5.0)
1 4.3 (1.1) 4.7 (2.0) 3.4 (2.3) 9.3 (2.0)
5 5.9 (0.8) 4.8 (2.0) 3.9 (1.5) 8.3 (1.5)
10 13.2 (1.0) 5.1 (2.0) 8.1 (1.0) 14.7 (1.0)
0b 3.1 (1.1) 3.1 (1.1) 7.5 (0.4) 7.5 (0.4)
0.1 5.7 (2.0) 0.8 (0.5) 2.8 (1.0) 13.9 (6.0)
1.5 0.5 5.0 (2.0) 5.7 (5.1) 4.3 (2.0) 11.7 (5.0)
1 4.3 (1.1) 4.7 (2.0) 4.7 (2.3) 9.3 (2.0)
5 6.2 (0.8) 5.9 (2.0) 5.3 (1.5) 9.8 (1.5)
10 13.5 (1.0) 6.1 (2.0) 8.1 (1.0) 17.4 (1.0)
0b 3.8 (1.1) 3.8 (1.1) 7.3 (0.4) 7.3 (0.4)
0.1 5.7 (2.0) 0.9 (0.5) 3.0 (1.0) 15.1 (6.0)
2.0 0.5 5.0 (2.0) 6.4 (5.1) 4.9 (2.0) 13.3 (5.0)
1 4.3 (1.1) 4.7 (2.0) 4.7 (2.3) 9.3 (2.0)
5 6.2 (0.8) 6.9 (2.0) 6.1 (1.5) 12.1 (1.5)
10 13.5 (1.0) 6.9 (2.0) 8.1 (1.0) 18.9 (1.0)
0b 3.8 (1.1) 3.8 (1.1) 7.3 (0.4) 7.3 (0.4)
0.1 5.7 (2.0) 0.9 (0.5) 3.9 (1.0) 15.5 (6.0)
2.5 0.5 5.0 (2.0) 7.2 (5.1) 5.6 (2.0) 15.5 (5.0)
1 4.3 (1.1) 4.7 (2.0) 4.7 (2.3) 9.3 (2.0)
5 6.2 (0.8) 6.9 (2.0) 6.6 (1.5) 14.8 (1.5)
10 13.5 (1.0) 6.9 (2.0) 8.1 (1.0) 18.4 (1.0)
0b 3.9 (1.1) 3.9 (1.1) 7.4 (0.4) 7.4 (0.4)
0.1 5.7 (2.0) 0.9 (0.5) 4.0 (1.0) 15.8 (6.0)
3.0 0.5 5.0 (2.0) 7.2 (5.1) 5.6 (2.0) 15.8 (5.0)
1 4.3 (1.1) 4.7 (2.0) 4.7 (2.3) 9.3 (2.0)
5 6.2 (0.8) 6.9 (2.0) 7.3 (1.5) 14.4 (1.5)
10 13.5 (1.0) 6.9 (2.0) 8.1 (1.0) 18.9 (1.0)
Key:-aStandard error values in parenthesis, b0 % surfactant preparations contained no surfactant

Acute ocular irritation results
All the test animals appeared healthy and were active. The triplicate
animals eye testing produced no ocular irritation with propylene
glycol (10 %) and 7 ointments (namely, four neat bases: EO, SO, HS,
NS; and chloramphenicol ointment prepared in SO, HW, and NS). The
other test items caused irritation to different measures: Two of 3
animals treated with neat HW showed corneal opacity, observed 1 h
after instillation, and partial conjunctiva swelling occurred in 1 of
the 2 positive corneal opacity cases. One of 3 animals showed
conjunctiva redness within 1 h of administering polysorbate 80 (10
%), chloramphenicol in HS and chloramphenicol in EO ointments.
The drug in EO ointment also produced mild swelling of the eye lid.
However, in every case, all the irritation signs were absent at the 24
h assessment. Apart from the eye irritation results noted above,
there were no other signs of gross toxicity, adverse pharmacologic
effects or abnormal behavior. Table 5 shows the individual total and
group mean ocular irritation scores. The maximum mean total score
of 0.67 was obtained for polysorbate 80 (10 %) solution and for
chloramphenicol in HS ointment; the drug in EO gave MMTS of 1.33,
while neat HW produced the highest score of 14.0. According to the
Kay and Calandra classification system, all the test items proved
practically non-irritant except neat HW that was classified as mildly
irritant.

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Table 5: Individual total scores and group mean scores for ocular Irritation
Test item/Rabbit ID/No; Sex (M/F) Time-interval of observation/Individual total score
Polysorbate 80 solution (10 %w/v) 1h 24 h 48 h 72 h
0061; F 2 0 0 0
0095; F 0 0 0 0
0023; F 0 0 0 0
Group Total 2 0 0 0
Group Mean Score 0.67 0 0 0
Chloramphenicol (1 %w/w) in Hydrous Sheabutter 1h 24 h 48 h 72 h
0161; F 0 0 0 0
0158; M 2 0 0 0
0164; F 0 0 0 0
Group Total 2 0 0 0
Group Mean Score 0.67 0 0 0
Chloramphenicol (1 %w/w) in Eye ointment basis 1h 24 h 48 h 72 h
0141; F 4 0 0 0
0088; F 0 0 0 0
0133; M 0 0 0 0
Group Total 4 0 0 0
Group Mean Score 1.33 0 0 0
Hydrous Wool Fat (neat) 1h 24 h 48 h 72 h
0067; M 0 0 0 0
0052; F 20 0 0 0
0033; M 22 0 0 0
Group Total 42 0 0 0
Group Mean Score 14 0 0 0

DISCUSSION
This study meets the need to conduct ocular tolerance testing of eye
ointment basis BP in order to investigate and validate its general
acceptance as the standard ophthalmic ointment vehicle. Erstwhile
untested probability that selected dermal ointment vehicles may be
suitable for ophthalmic drug delivery has also been elucidated. The
corneal epithelium is more permeable to medicinal agents than skin
epithelium [27]. The in vivo rabbit eye test approximates to human
physiological and anatomical characteristics and response [28],
hence it was appropriate for the study.
The drug release mechanism of chloramphenicol from the bases was
determined as Higuchi (diffusion-controlled) model. Similar earlier
studies reported fluconazole to be released from water-soluble
ointment bases following zero order kinetics [29] and metronidazole
released from different ointment base types following the Higuchi
kinetics [19]. The selective enhancement of release of
chloramphenicol from the EO and SO formulations is not unusual.
Earlier studies indicated that inclusion of surfactant in semisolid
drug delivery vehicle may sometimes increase [30] or decrease the
release of incorporated drug [31, 32]. Characteristics demonstrated
by neat HW base in this study were unique and different from those
of other bases namely, non-conformity of its drug release potential
with the observed trends against WAC and temperature
susceptibility, and its (mild) ocular irritancy potential.
CONCLUSION
Chloramphenicol eye ointment formulations in five different bases,
with and without surfactant, investigated in this study were all well
tolerated and non-irritant to the eye. One test item only (neat HW)
gave an overall eye irritation score (MMTS) of mild irritancy. All the
four neat bases investigated in parallel as alternatives to EO showed
greater propensity for delivering the drug from ointment than EO,
with NS and HS demonstrating the highest potentials for both the
rate and extent of drug release. WAC and softening temperature of
base apparently influenced its drug release propensity. Whereas the
presence of polysorbate 80 in chloramphenicol-in-EO formulation
did not appreciably improve the drug release from the base,
propylene glycol (10 %w/v) in similar formulation caused more
than three-fold enhancement of the drug release.
ACKNOWLEDGEMENT
The authors thankfully acknowledge: Dr. Adesoji Matthew Olaniyan
for providing freshly processed sheabutter at NCAM, Ilorin Nigeria,
for this work; and the veterinary expertise of Mr. Olumide Oyeniyi
Kolawole of Biological Gardens Unit, Zoology Department, OAU, Ile-
Ife in the procurement and maintenance of the experimental
animals; also Mr. Oladapo Kolapo Adeboye and Miss Olaitan
Oluwayemisi Abiona for their laboratory assistance with the animal
tests.
CONFLICT OF INTERESTS
All authors have none to declare
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